1. Biochemical Basis for Increased Activity of Ebola Glycoprotein in the 2013–16 Epidemic 
May K. Wang, Sun-Young Lim, Soo Mi Lee, James M. Cunningham 
Cell Host & Microbe 
Volume 21, Issue 3, Pages (March 2017)
DOI: /j.chom
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2. Cell Host & Microbe 2017 21, 367-375DOI: (10.1016/j.chom.2017.02.002)
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3. Figure 1
Sensitivity of Makona Virus Transduction to the NPC1-Targeting Inhibitor 3.47
(A and B) Vero cells were cultured in media containing increasing concentrations of 3.47 (0–2 μM) for 1 hr before transduction with MLV particles encoding GFP and pseudotyped with either FL GP from Makona, Makona A82V or Mayinga viruses (A), or mucin-like domain deleted GP from Makona or Makona A82V viruses (B). Virus transduction is reported as the percentage of GFP-positive cells relative to cells exposed to DMSO alone. Data are mean ± SD (n = 3). See also Table 1. Data in Figure 1B was collected in the same experiment as data in Figures 2B, 3B, and S4 and Table 1.
Cell Host & Microbe  , DOI: ( /j.chom )
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4. Figure 2 Makona A82V GP Has Increased NPC1-Induced Specific Activity
(A) Lysosome membranes from CHONPC1 and CHONull cells were incubated with increasing concentrations of Makona or Makona A82V GPΔTM that were cleaved to expose the NPC1 binding domain (top) or incubated with increasing concentrations of 3.47 for 30 min prior to the addition of cleaved Makona or Makona A82V GPΔTM (bottom). Membrane-bound and input GP were determined by immunoblot using anti-GP1 serum. See also Figure S3.
(B) Vero cells were grown in media containing the indicated concentrations of 3.47 (0–2 μM) for 1 hr and then transduced with MLV particles encoding GFP and pseudotyped with Makona or Makona A82V GP. Virus transduction is reported as the percentage of GFP-positive cells relative to cells exposed to DMSO vehicle alone. Data are mean ± SD (n = 3). See also Table 1. Data were collected in the same experiment as data in Figures 1B, 3B, and S4 and Table 1.
(C) Comparison of Mayinga and Mayinga A82V GPΔTM binding to lysosome membranes alone (top) or in the presence of 3.47 (bottom) was performed as in (A). See also Figure S3.
Cell Host & Microbe  , DOI: ( /j.chom )
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5. Figure 3
Residue 544 in GP2 Is Also a Determinant of GP Specific Activity and Transduction
(A) Lysosome membranes from CHONPC1 and CHONull cells were incubated with increasing concentrations of cleaved Mayinga or Makona GPΔTM (top) or incubated with increasing concentrations of 3.47 for 30 min prior to the addition of cleaved Mayinga or Makona GPΔTM (bottom). Membrane-bound and input GP1 were analyzed by immunoblot using anti-GP1 serum. See also Figure S3.
(B) Vero cells were cultured in media containing the indicated concentrations of 3.47 (0–2 μM) for 1 hr and then challenged with MLV particles encoding GFP and pseudotyped with Mayinga or Mayinga I544T GP. Virus transduction is reported as the percentage of GFP-positive cells relative to cells exposed to DMSO vehicle alone. Data are mean ± SD (n = 3). See also Table 1. Data were collected in the same experiment as data in Figures 1B, 2B, and S4 and Table 1.
(C) Comparison of Mayinga and Mayinga I544T GPΔTM binding to lysosome membranes alone (top) or in the presence of 3.47 (bottom) were performed as in (A). See also Figure S3.
Cell Host & Microbe  , DOI: ( /j.chom )
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6. Figure 4
Comparative Analysis of Receptor Binding to Makona and Mayinga GPs
(A) MLV particles pseudotyped with the indicated GPs were cleaved to expose the NPC1 binding domain and bound to plates using KZ52 mAb. Bound particles were incubated with increasing concentrations of purified NPC1 Domain C-His6. Bound NPC1 Domain C was detected using α-His6 antibody. Binding is reported as mean ± SD (n = 4) of the percent of maximum binding for each GP. Pairwise comparison indicates that the affinities of these GPs for NPC1 Domain C are not statistically significant (p = 0.18). See also Figure S3.
(B) MLV particles pseudotyped with the indicated GPs were cleaved and then heated for 30 min at the specified temperatures (T = 25°C–51.7°C). After cooling to room temperature, virus particles were captured on ELISA plates coated with the GP2 prefusion conformation-specific mAb KZ52. Results are mean ± SD (n = 3) of the percent of binding for each GP at T = 25°C. Makona versus Makona A82V: p < ; Mayinga versus Mayinga A82V: p < ; Mayinga versus Mayinga I544T: p < ; Mayinga versus Makona A82V: p = 0.4; Makona versus Mayinga I544T: p = 1. See also Figure S5.
Cell Host & Microbe  , DOI: ( /j.chom )
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7. Figure 5 Functional Comparison of Makona and Mayinga GPs
(A) Analysis of requirements for the host factor cathepsin B. Vero cells were grown in media containing the cathepsin B inhibitor CA074 (10 μM) for 4 hr before transduction with MLV particles encoding GFP and pseudotyped with the indicated GP. Virus transduction is reported as the percentage of GFP-positive cells relative to cells exposed to DMSO vehicle alone. Data are mean ± SD (n = 3). ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < Mayinga versus Mayinga I544T: n.s.; Makona versus Mayinga I544T: n.s.; Mayinga A82V versus Mayinga A82V/A503V: n.s.
(B) Stability of Makona and Mayinga GPs on virus particles. MLV particles encoding GFP and pseudotyped with the indicated GP were incubated at 37°C for the specified times (T = 0–76 hr) and then assayed for transduction of Vero cells. Data are mean ± SD (n = 3) and reported as the percentage of infection at T0. Makona versus Makona A82V: p = 0.02; Mayinga versus Mayinga A82V: p = 0.04; Mayinga versus Mayinga I544T: p = 0.03; Mayinga versus Makona A82V: p = 0.8; Makona versus Mayinga I544T: p = 1.
Cell Host & Microbe  , DOI: ( /j.chom )
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